EP0948527B1 - Cyclosporin derivatives, their preparation and pharmaceutical compositions containing them - Google Patents

Abstract

A cyclosporin compound of the formula (I): wherein Alk and R are as defined herein, or a pharmaceutically acceptable salt thereof, which derivatives are is useful in the treatment and/or prophylaxis of retrovirus infections.

Description

leurs sels, leur préparation et les compositions pharmaceutiques qui les contiennent.The present invention relates to cyclosporine derivatives of general formula:

their salts, their preparation and the pharmaceutical compositions containing them.

These derivatives are useful for the treatment and / or prophylaxis of retrovirus infections, especially AIDS (syndrome acquired immunodeficiency) and associated syndromes [ARC (AIDS related complex)]. They have the advantage of being very weak immunosuppressants.

Cyclosporine derivatives modified at position -3 have been previously described as immunosuppressive agents, in the patent European EP 194972.

Cyclosporine [4'-hydroxy-Meleu] ⁴ derivatives have been described in European patent EP 484 281. These derivatives are useful for the treatment of AIDS and are not immunosuppressive.

• Alk représente un radical alcoylène contenant 2 à 6 atomes de carbone en chaíne droite ou ramifiée ou cycloalcoyléne contenant 3 à 6 atomes de carbone et
• R représente
  • soit un radical carboxy ou alcoyloxycarbonyle,
  • soit un radical -NR₁R₂ pour lequel R₁ et R₂ identiques ou différents représentent des atomes d'hydrogène, ou des radicaux alcoyle, alcènyle (2 à 4C), cycloalcoyle (3 à 6C), phényle éventuellement substitué (par un atome d'halogène, alcoyloxy, alcoyloxycarbonyle, amino, alcoylamino ou dialcoylamino). ou représentent des radicaux benzyle ou hétérocyclyle saturé ou insaturé contenant 5 ou 6 chaínons et 1 à 3 hétéroatomes, ou pour lequel R₁ et R₂ forment avec l'atome d'azote auquel ils sont rattachés un hétérocycle contenant 4 à 6 chaínons, saturé ou insaturé, pouvant contenir un autre hétéroatome choisi parmi l'azote, l'oxygène ou le soufre et éventuellement substitué par alcoyle, phényle ou benzyle,
  • soit un radical de formule générale :
    
    pour lequel R₁ et R₂ sont définis comme ci-dessus, R₃ représente un atome d'hydrogène ou un radical alcoyle et n est un nombre entier de 2 à 4,

les portions ou radicaux alcoyle définis ci-dessus étant droits ou ramifiés et contenant 1 à 4 atomes de carbone,
ainsi que leurs sels pharmaceutiquement acceptables lorsqu'ils existent, sont particulièrement intéressants du fait de leur activité puissante et de leur caractère très faiblement immunosuppresseur.It has now been found that the cyclosporine derivatives of general formula (I) in which:

• Alk represents an alkylene radical containing 2 to 6 carbon atoms in a straight or branched chain or cycloalkylene containing 3 to 6 carbon atoms and
• R represents
  • either a carboxy or alkyloxycarbonyl radical,
  • either a radical -NR ₁ R ₂ for which R ₁ and R ₂ identical or different represent hydrogen atoms, or alkyl, alkenyl (2 to 4C), cycloalkyl (3 to 6C), phenyl radicals optionally substituted (by a halogen atom, alkyloxy, alkyloxycarbonyl, amino, alkyllamino or dialcoylamino). or represent saturated or unsaturated benzyl or heterocyclyl radicals containing 5 or 6 chains and 1 to 3 heteroatoms, or for which R ₁ and R ₂ form, with the nitrogen atom to which they are attached, a heterocycle containing 4 to 6 chains, saturated or unsaturated, which may contain another heteroatom chosen from nitrogen, oxygen or sulfur and optionally substituted by alkyl, phenyl or benzyl,
  • either a radical of general formula:
    
    for which R ₁ and R ₂ are defined as above, R ₃ represents a hydrogen atom or an alkyl radical and n is an integer from 2 to 4,

the alkyl portions or radicals defined above being straight or branched and containing 1 to 4 carbon atoms,
as well as their pharmaceutically acceptable salts when they exist, are particularly advantageous because of their potent activity and their very weakly immunosuppressive character.

In the general formula (I), when R ₁ and / or R ₂ represent heterocyclyl, it can be advantageously chosen from pyridyl, tetrahydropyridyl, piperidyl, imidazolyl, oxazolyl, thiazolyl.

When R ₁ and R ₂ form heterocyclyl with the nitrogen atom to which they are attached, for example, the heterocyclyl radical can be chosen from azetidinyl, piperidyl, piperazinyl, N-methyl piperazinyl, N-phenyl piperazinyl, N -benzyl piperazinyl, pyridyl, imidazolyl, morpholino, thiomorpholino, tetrahydropyridyl, methyl tetrahydropyridyl (for example 4-methyl tetrahydropyridyl), phenyl tetrahydropyridyl (for example 4-phenyl tetrahydropyridyl).

According to the present invention, the products of general formula (I) can be obtained by the action of a disulfide of general formula: R-Alk-SS-Alk-R in which R and Alk are defined as above, and the functions of which which may interfere with the reaction, where appropriate, have been protected beforehand, on an activated form of cyclosporin A, then if necessary eliminates the protective radical.

By activated form of cyclosporin A is meant an activated form on sarcosine in position 3. This activated form of cyclosporine A is preferably prepared in situ. activation generally takes place under an inert atmosphere, by treatment with an organometallic derivative (notably lithian, such as n-butyllithium, lithium diisopropyl amide or a mixture by example). It is also possible to prepare the activated form of cyclosporine A in liquid ammonia in the presence of an amide alkaline (sodium, lithium for example), at a temperature between -32 and -38 ° C, in an ether (in particular tetrahydrofuran, t.butylethyl ether or a mixture).

The disulfide of general formula (II) is added advantageously in an organic solvent such as a hydrocarbon (hexane for example) or an ether (diethyl ether, tetrahydrofuran or t.butylmethylether for example) at a temperature between -78 and 0 ° C. It is sometimes preferable to operate under nitrogen.

When the substituents of the radical R can interfere with the reaction, it is best to protect them beforehand with compatible radicals that can be set up and removed without touch the rest of the molecule. In addition, the hydroxy radical present on cyclosporine can be optionally protected by any group which does not interfere with the reaction.

By way of example, the protective groups can be chosen among the radicals described by T.W. GREENE, Protective Groups in Organic Synthesis, J. Wiley-Interscience Publication (1991) or by Mc Omie, Protective Groups in Organic Chemistry, Plenum Press (1973).

The disulfide of general formula (II) can be prepared according to or by analogy with the methods described in the examples.

The new cyclosporine derivatives of general formula (I) can be purified if necessary by physical methods such as crystallization or chromatography.

Cyclosporine derivatives according to the invention for which R is carboxy can be transformed into metal salts or salts addition with a nitrogenous base according to methods known per se. These salts can be obtained by the action of a metal base (by alkaline or alkaline earth), ammonia or a amine on a product according to the invention, in a suitable solvent such as water or alcohol. The salt formed precipitates after possible concentration of the solution; it is separated by filtration.

As examples of pharmaceutically acceptable salts can be cited the salts with alkali metals (sodium, potassium, lithium) or with alkaline earth metals (magnesium, calcium), salt ammonium, nitrogen base salts (ethanolamine, diethanolamine, trimethylamine, triethylamine, methylamine, propylamine, diisopropylamine, N, N-dimethylethanolamine, benzylamine, dicyclohexylamine, N-benzylphenethylamine, N, N'-dibenzylethylenediamine, diphenylenediamine, benzhydrylamine, quinine, choline, arginine, lysine, leucine, dibenzylamine).

The cyclosporine derivatives according to the invention for which R is NR ₁ R ₂ can be converted into addition salts with acids, by known methods. It is understood that these salts also come within the scope of the present invention.

As examples of addition salts with pharmaceutically acids acceptable, may be mentioned the salts formed with acids minerals (hydrochlorides, hydrobromides, sulfates, nitrates, phosphates) or with organic acids (succinates, fumarates, tartrates, acetates, propionates, maleates, citrates, methanesulfonates, ethanesulfonates, p.toluenesulfonates, isethionates, embonates or with substitution derivatives of these compounds).

The new cyclosporine derivatives according to the present invention are particularly useful for prophylaxis and treatment retrovirus diseases and more particularly AIDS and associated syndromes. By prophylaxis is meant in particular the treatment of subjects who have been exposed to HIV viruses, especially asymptomatic HIV-positive people who have the risk of developing the disease in the coming months or years after primary infection.

The products according to the invention exhibit anti-retrovirus activity at concentrations without cytotoxic effect or cytostatic.

The activity of the products of general formula (I) has been demonstrated in the techniques described by Pauwells et al., J. Virol. Meth., 20 , 309 (1988) and by O. Schwatz et al., AIDS Research and Human Retroviruses, 4 (6) , 441-48 (1988) and cited by JF Mayaux et al. Proc. Nat. Acad. Sci. USA, 91, 3564-68 (1994). In these techniques, the products according to the invention have been shown to be active at concentrations of 3 to 350 nM (IC ₅₀ ).

• Alk représente un radical alcoyléne contenant 2 à 6 atomes de carbone en chaíne droite ou ramifiée et
• R représente un radical -NR₁R₂ pour lequel R₁ et R₂ identiques ou différents représentent des atomes d'hydrogène, ou des radicaux alcoyle, alcènyle (2 à 4C), phényle éventuellement substitué (par un atome d'halogène, alcoyloxy, alcoyloxycarbonyle, amino, alcoylamino ou dialcoylamino), ou représentent des radicaux benzyle ou pour lequel R₁ et R₂ forment avec l'atome d'azote auquel ils sont rattachés un hétérocycle contenant 4 à 6 chaínons, saturé ou insaturé, pouvant contenir un autre hétéroatome choisi parmi l'azote, l'oxygène ou le soufre et éventuellement substitué par alcoyle,

les portions ou radicaux alcoyle définis ci-dessus étant droits ou ramifiés et contenant 1 à 4 atomes de carbone, ainsi que leurs sels pharmaceutiquement acceptables lorsqu'ils existent ;Of particular interest are the products of general formula (I) for which:

• Alk represents an alkyl radical containing 2 to 6 carbon atoms in a straight or branched chain and
• R represents a radical -NR ₁ R ₂ for which R ₁ and R _{2, which are} identical or different, represent hydrogen atoms, or alkyl, alkenyl (2 to 4C), phenyl radicals optionally substituted (by a halogen atom, alkyloxy , alkyloxycarbonyl, amino, alkyllamino or dialkoylamino), or represent benzyl radicals or for which R ₁ and R ₂ form with the nitrogen atom to which they are attached a heterocycle containing 4 to 6 chains, saturated or unsaturated, which may contain a another heteroatom chosen from nitrogen, oxygen or sulfur and optionally substituted with alkyl,

the alkyl portions or radicals defined above being straight or branched and containing 1 to 4 carbon atoms, as well as their pharmaceutically acceptable salts when they exist;

• Alk représente un radical alcoyiène contenant 2 à 5 atomes de carbone en chaíne droite ou ramifiée et
• R représente un radical -NR₁R₂ pour lequel R₁ et R₂ identiques ou différents représentent des atomes d'hydrogène, ou des radicaux alcoyle, allyle, phényle ou benzyle ou pour lequel R₁ et R₂ forment avec l'atome d'azote auquel ils sont rattachés un hétérocycle choisi parmi azétidinyle, pipéridyle, pipérazinyle, N-méthyl pipérazinyle, N-phényl pipérazinyle, N-benzyl pipérazinyle, imidazolyle, morpholino, tétrahydropyridyle, méthyl tétrahydropyridyle, phényl tétrahydropyridyle.

les portions ou radicaux alcoyle définis ci-dessus étant droits ou ramifiés et contenant 1 à 4 atomes de carbone, ainsi que leurs sels pharmaceutiquement acceptables lorsqu'ils existent ;According to an even more preferred aspect of the invention, the cyclosporine derivatives of general formula (I) for which:

• Alk represents an alkylene radical containing 2 to 5 carbon atoms in a straight or branched chain and
• R represents a radical -NR ₁ R ₂ for which R ₁ and R ₂ identical or different represent hydrogen atoms, or alkyl, allyl, phenyl or benzyl radicals or for which R ₁ and R ₂ form with the atom d nitrogen to which they are attached a heterocycle chosen from azetidinyl, piperidyl, piperazinyl, N-methyl piperazinyl, N-phenyl piperazinyl, N-benzyl piperazinyl, imidazolyl, morpholino, tetrahydropyridyl, methyl tetrahydropyridyl, phenyl tetrahyd.

the alkyl portions or radicals defined above being straight or branched and containing 1 to 4 carbon atoms, as well as their pharmaceutically acceptable salts when they exist;

• [(R)-2-(N,N-diéthylamino)éthylthio-Sar]³-cyclosporine A ;
• [(R)-2-(N,N-diméthylamino)éthylthio-Sar]³-cyclosporine A ;
• [(R)-2-(1-pipéridyl)-éthylthio-Sar]³ cyclosporine A ;
• [(R)-2-(N-méthyl-N-i.propylamino)éthylthio-Sar]³ cyclosporine A ;
• [(R)-2-(N-méthyl-N-t.butylamino)éthylthio-Sar]³ cyclosporine A ;

ainsi que leurs sels pharmaceutiquement acceptables lorsqu'ils existent.And among these products in particular the following cyclosporine derivatives:

• [(R) -2- (N, N-diethylamino) ethylthio-Sar] ³ -cyclosporin A;
• [(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ -cyclosporin A;
• [(R) -2- (1-piperidyl) -ethylthio-Sar] ³ cyclosporin A;
• [(R) -2- (N-methyl-Ni.propylamino) ethylthio-Sar] ³ cyclosporine A;
• [(R) -2- (N-methyl-Nt.butylamino) ethylthio-Sar] ³ cyclosporin A;

as well as their pharmaceutically acceptable salts when they exist.

The following examples, given without limitation, illustrate the present invention.

Example 1

[(R) -2- (N, N-diethylamino) ethylthio-Sar] ^3- cyclosporin A methane sulfonate is prepared according to the following method:

To a solution of 3.53 cm ³ of diisopropylamine (previously distilled on calcium hydride) in 45 cm ³ of tetrahydrofuran (previously distilled on sodium), cooled to a temperature in the region of -10 ° C and under nitrogen, the mixture is added in 20 minutes, 15.6 cm ³ of a 1.6 M solution of n-butyllithium in hexane while maintaining the temperature at 0 ° C. The mixture is stirred at 0 ° C for 20 minutes, then it is cooled to a temperature in the region of -78 ° C. The solution thus obtained is transferred, under nitrogen, by a transfer rod, to a solution of 2 g of cyclosporine A in 40 cm ³ of tetrahydrofuran previously cooled to a temperature close to -78 ° C while maintaining the temperature around -75 ° vs. The resulting mixture is stirred at -75 ° C for 10 minutes, then 6.3 cm ³ of a solution of 1.6 M n-butyllithium in hexane are added over 4 minutes. Stirring is continued for 20 minutes then 8.8 g of di- [2- (N, N-diethylamino) ethyl] disulfide are added over 2 minutes while maintaining the temperature around -75 ° C. The mixture is stirred at a temperature in the region of -75 ° C for 30 minutes and then at 0 ° C for 18 hours. A mixture of 50 cm ³ of ice-cold distilled water and 36% aqueous hydrochloric acid is poured onto the reaction mixture to obtain a pH close to 7, then the mixture is decanted, the aqueous phase washed with 30 cm ³ of diethyl ether. The organic extracts are combined, washed with 50 cm ³ of saturated sodium chloride solution, dried over sodium sulfate, filtered and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The solid obtained is purified by 2 successive flash chromatographies on a silica column (0.04-0.063 mm) (eluent methanol-water-dichloromethane 14: 2: 84 by volume then dichloromethane-methanol 93: 7 by volume), collecting fractions of 10 cm ³ . The fractions containing the expected product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to give 0.245 g of [(R) -2- (N, N-diethylamino) ethylthio-Sar] ³ cyclosporine A as a white solid.

0.245 g of [(R) -2- (N, N-diethylamino) ethylthio-Sar] ^3- cyclosporin A in solution in 15 cm ³ of diethyl ether is placed in a flask with stirring. After filtration, 15 mg of methanesulfonic acid dissolved in 2 cm ³ of diethyl ether are added over 30 seconds. The white precipitate obtained is filtered, washed with 3 times 2 cm ³ of diethyl ether and then with 2 cm ³ of pentane. 0.067 g of [(R) -2 (N, N-diethylamino) ethylthio-Sar] ³ cyclosporin A methane sulfonate is thus obtained in the form of a white solid, melting at 155 ° C (dec.).

1 H NMR spectrum (400 MHz, CDCl ₃ , δ in ppm); 1.24 (d, J = 7 Hz, 3H: CH ₃ 8β); 1.31 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 1.31 (t, J = 7 Hz, 6H: CH ₃ of ethyl 2-diethylaminoethylthio in 3α); 1.60 (d, J = 5 Hz, 3H: CH ₃ in 1γ); 2.38 (mt, 1H: CH 5β); 2.66 - 2.75 -3.08 - 3.10 - 3.22 - 3.41 and 3.46 (7 s; respectively 6H - 3H - 3H - 3H - 3H - 3H and 3H: 7 NCH ₃ and CH ₃ methanesulfonate); from 2.90 to 3.35 (mf, 8H: SCH ₂ CH ₂ N of 2-diethylaminoethylthio in 3α and N CH ₂ of ethyl of 2-diethylaminoethylthio in 3α); 3.76 (mt, 1H: CH 1β); 4.48 (mt, 1H: CH 7α); 4.64 (t, J = 9 Hz, 1H: CH 5α); 4.81 (mt, 1H: CH 8α); from 4.95 to 5.05 (mt, 2H: CH 2 α and CH α of a leucine); 5.06 (d, J = 11 Hz, 1H: CH 11α); 5.18 (dd, J = 12 and 4 Hz, 1H: CH α of a leucine); 5.20 to 5.35 (mt, 2H: CH = CH); 5.41 (d, J = 6 Hz, 1H: CH 1α); 5.67 (dd, J = 10 and 4 Hz, 1H: CH α of a leucine); 5.90 (s, 1H: CH 3α); 7.17 (d, J = 9 Hz, 1H: CONH in 5); 7.39 (d, J = 8 Hz, 1H: CONH in 8); 7.68 (d, J = 7.5 Hz, 1H: CONH at 7); 8.04 (d, J = 9.5 Hz, 1H: CONH in 2).

Di- [2- (N, N-ethylamino) ethyl] disulfide can be prepared according to the method described in H. Gilman J. Am. Chem. Soc. (1945) 67 , 1846

Example 2

[(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ -cyclosporin A is prepared by the following method:

To a solution of 26.5 cm ³ of diisopropylamine (previously distilled on calcium hydride) in 300 cm ³ of tetrahydrofuran (previously distilled on sodium) cooled to a temperature in the region of -5 ° C and under nitrogen, added in 30 minutes , 117 cm ³ of a 1.6 M solution of n-butyllithium in hexane while maintaining the temperature at 0 ° C. The mixture is stirred at 0 ° C for 20 minutes, then it is cooled to a temperature in the region of -78 ° C. The solution thus obtained is transferred, under nitrogen, by a transfer rod, to a solution of 15 g of cyclosporin A in 270 cm ³ of tetrahydrofuran previously cooled to a temperature close to -76 ° C while maintaining the temperature around -70 ° vs. The resulting mixture is stirred at -78 ° C for 10 minutes, then 47 cm ³ of a solution of 1.6 M n-butyllithium in hexane are added over 10 minutes. Stirring is continued for 5 minutes and then 52 g of di- [2- (N, N-dimethylamino) ethyl] disulfide are added slowly while maintaining the temperature around -75 ° C. The mixture is stirred at a temperature in the region of -78 ° C for 30 minutes and then at 0 ° C for 18 hours. 120 cm ³ of ice-cold distilled water added with 80 cm ³ of 12N hydrochloric acid are poured onto the reaction mixture, kept stirring at -20 ° C., so as to obtain a pH close to 7, then the mixture is decanted and the phase aqueous washed with 100 cm ³ of ethyl acetate. The organic extracts are combined, washed with a saturated sodium chloride solution, dried over sodium sulfate, filtered and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The solid obtained is dissolved in 200 cm ³ of toluene and 1000 cm ³ of distilled water added with 12N hydrochloric acid to bring the pH to 2. The phases are decanted. The aqueous phase is washed back with 100 cm ³ of toluene and the toluene phases combined. These are washed with 200 cm ³ of distilled water, acidified to a pH close to 3. The aqueous phases are combined, added with 200 cm ³ of toluene and then neutralized with an aqueous solution of sodium bicarbonate. The organic phase is decanted, the aqueous phase washed with 100 cm ³ of toluene. The organic phases are combined, dried over sodium sulfate, filtered and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to give a solid which is purified by flash chromatography on a silica column (0.04 -0.063 mm) (eluent dichloromethane-methanol 93.5: 6.5 by volume) and collecting 35 cm ³ fractions. The fractions containing the expected product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to give a solid which is concreted in 30 cm ³ of pentane. After filtration, 1.94 g of [(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ cyclosporin A are obtained in the form of a white solid, melting at around 140 ° C.

¹ H NMR spectrum (400 MHz, CDCl ₃ , δ in ppm): 1.28 (d, J = 7 Hz, 3H: CH ₃ 8β); 1.37 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 1.63 (mt, 3H: CH ₃ in 1γ); 2.25 (s, 6H: N (CH ₃ ) ₂ of 2-dimethylaminoethylthio in 3α); 2.40 (mt, 1H: CH 5β); 2.50 to 2.80 (mf, 4H: SCH ₂ CH ₂ N of 2-dimethylaminoethylthio in 3α); 2.71 - 3.13 - 3.14 - 3.28 - 3.46 and 3.52 (6 s; 6H - 3H - 3H - 3H - 3H and 3H, respectively: 7 NCH ₃ ); 3.65 (d, J = 6 Hz, 1H: OH in 1β); 3.78 (mt, 1H: CH 1β); 4.56 (mt, 1H: CH 7α); 4.67 (t, J = 9 Hz, 1H: CH 5α); 4.85 (mt, 1H: CH 8α); 4.99 (dd, J = 9 and 6 Hz, 1H: CH α of a leucine); 5.00 to 5.15 (mt, 2H: CH 2α and CH α of a leucine); 5.15 (d, J = 11 Hz, 1H: CH 11α); 5.25 (dd, J = 12 and 4 Hz, 1H: CH α of a leucine); 5.30 to 5.45 (mt, 2H: CH = CH); 5.51 (d, J = 6 Hz, 1H: CH 1α); 5.72 (dd, J = 10.5 and 4 Hz, 1H: CH α of a leucine); 6.02 (s, 1H: CH 3α); 7.18 (d, J = 8 Hz, 1H: CONH at 8); 7.35 (d, J = 9 Hz, 1H: CONH in 5); 7.68 (d, J = 7.5 Hz, 1H: CONH at 7); 7.96 (d, J = 9.5 Hz, 1H: CONH in 2).

The methane sulfonate salt of [(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ cyclosporine A, is prepared as follows: 360 mg of [(R) is placed in a flask -2- (N, N-dimethylamino) ethylthio-Sar] ³ cyclosporin A dissolved in 6 cm ³ of diethyl ether then 24 mg of methanesulfonic acid dissolved in 1 cm ³ of diethyl ether are added in 30 seconds. The white precipitate obtained is filtered, washed with 3 times 2 cm ³ of diethyl ether and then with 5 cm ³ of pentane. After drying at 50 ° C. under reduced pressure (40 kPa), 0.328 mg of [(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ cyclosporin A methane sulfonate is thus obtained in the form of a white solid melting at 155 ° C (dec.).

¹ H NMR spectrum (400 MHz, (CD ₃ ) ₂ SO d6, δ in ppm): 1.21 (d, J = 7.5 Hz, 3H: CH ₃ 8β); 1.29 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 1.69 (d, J = 6.5 Hz, 3H: CH ₃ in 1γ); 1.99 (mt, 1H: CH 5β); 2.35 (s, 3H: CH ₃ of methanesulfonate); from 2.45 to 2.70 (mt, 2H: SCH ₂ of 2-dimethylaminoethylthio in 3α); 2.64 - 2.80 - 2.86 - 2.93 - 2.99 and 3.17 (6 s, respectively 3H - 6H - 9H - 3H - 3H and 3H: 7 NCH ₃ and NCH ₃ of 2-dimethylaminoethylthio in 3α); from 3.25 to 3.40 (mf, 2H: CH ₂ N of 2-dimethylaminoethylthio in 3α); 3.99 (mt. 1H: CH 1β); 4.15 (mt, 1H: CH 7α); 4.26 (t, J = 9 Hz, 1H: CH 5α); 4.42 (broad s, 1H: OH in 1β); 4.79 (mt, 1H: CH 8α); 4.89 (mt, 1H: CH 2α); 5.00 to 5.15 (mt, 1H: CH α of a leucine); 5.11 (d, J = 11 Hz, 1H: CH 11α); 5.23 (mt, 2H: CH 1α and CH α of a leucine); 5.33 (dd, J = 10 and 5 Hz, 1H: CH α of a leucine); from 5.30 to 5.50 and 5.62 (2 mts, 1H each: CH = CH); 5.48 (dd, J = 11 and 5 Hz, 1H: CH α of a leucine); 6.87 (s, 1H: CH 3α); 7.64 (d, J = 7.5 Hz, 1H: CONH at 7); 8.24 (d, J = 9.5 Hz, 1H: CONH in 2); 8.28 (d, J = 8 Hz, 1H: CONH in 8); 8.68 (d, J = 9 Hz, 1H: CONH in 5); 9.28 (mf, 1H: SO ₃ H of methanesulfonate).

Example 3

[(R) -2- (1-piperidyl) -ethylthio-Sar] ³ cyclosporine A is prepared according to the following method:

To a solution of 7.0 cm ³ of diisopropylamine in 40 cm ³ of tetrahydrofuran cooled to a temperature in the region of 0 ° C. and under argon, 31 cm ³ of a solution of 1.6 n-butyllithium is added dropwise M in hexane. The mixture is stirred at 0 ° C for 20 minutes, then it is cooled to a temperature in the region of -78 ° C and a solution of 4.0 g of cyclosporin A and 6.0 cm ³ of 1,3-dimethyl- 3,4,5,6-tetrahydro-2 (1H) -pyrimidinone in 20 cm ³ of tetrahydrofuran under argon previously cooled to a temperature in the region of -78 ° C is added dropwise. The resulting mixture is stirred at a temperature in the region of -40 ° C for 20 minutes. then at a temperature in the region of -78 ° C. for 1 h, then a solution of 13.8 g of di- [2- (1-piperidyl) ethyl disulfide] in 20 cm ³ of tetrahydrofuran is added dropwise. The mixture is then stirred at a temperature in the region of -78 ° C for 10 minutes, then it is allowed to warm up to a temperature in the region of 20 ° C. It is then treated with 100 cm ³ of water. The aqueous phase is extracted 3 times with 100 cm ³ of ethyl acetate then the organic extracts are combined, washed 4 times with 50 cm ³ of water, dried over magnesium sulfate and concentrated under reduced pressure (2.7 kPa ) at a temperature close to 40 ° C. The oil obtained is chromatographed on a column containing 1.6 kg of silica (0.02-0.05 mm) eluted with ethyl acetate at atmospheric pressure and collecting 75 cm ³ fractions. The fractions containing the expected product are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The residue is dissolved in 20 cm ³ of ethyl acetate, filtered through paper, then reconcentrated under the same conditions. The residue obtained is taken up in 20 cm ³ of ethyl ether, reconcentrated in the same way, then dried to constant weight. 0.4 g of [(R) -2- (1-piperidyl) ethylthio-Sar] ³ cyclosporin A is thus obtained in the form of a straw yellow solid melting at 132 ° C.

¹ H NMR spectrum (400 MHz, CDCl ₃ , δ in ppm): 1.30 (d, J = 7 Hz, 3H: CH ₃ 8β); 1.37 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 1.65 (d, J = 5 Hz, 3H: CH ₃ in 1γ); from 1.90 to 3.10 (mt, 14H: CH ₂ of SCH ₂ CH ₂ Npiperidine in 3α); 2.47 (mt, 1H: CH 5β); 2.70 - 2.72 - 3.13 - 3.18 - 3.27 - 3.45 and 3.52 (7 s, 3H each: 7 NCH ₃ ); 3.77 (mt, 1H: CH 1β); 4.55 (mt, 1H: CH 7α); 4.65 (t, J = 9 Hz, 1H: CH 5α); 4.86 (mt, 1H: CH 8α) 4.99 (dd, J = 9 and 6 Hz, 1H: CH α of a leucine); 5.00 to 5.15 (mt, 2H: CH 2α and CH α of a leucine); 5.15 (d, J = 11 Hz, 1H: CH 11α); 5.24 (dd, J = 12 and 4 Hz, 1H: CH α of a leucine); of. 5.25-5.45 (mt, 2H: CH = CH); 5.50 (d, J = 6 Hz, 1H: CH 1α); 5.72 (dd, J = 10.5 and 4 Hz, 1H: CH α of a leucine); 6.22 (s, 1H: CH 3α); 7.18 (d, J = 8 Hz, 1H: CONH at 8); 7.38 (d, J = 9 Hz, 1H: CONH in 5); 7.70 (d, J = 7.5 Hz, 1H: CONH at 7); 7.95 (d, J = 9.5 Hz, 1H: CONH in 2).

Di- [2- (1-piperidyl) -ethyl] disulfide can be prepared according to the method described by RC Fuson in J. Org. Chem., 11 , 487 (1946).

Example 4

[(R) -2- (N-methyl-Ni.propylamino) ethylthio-Sar] ³ cyclosporin A is prepared according to the following method:

To 100 cm ³ of ammonia maintained at a temperature in the region of -33 ° C., 100 mg of metallic sodium is added, then 100 mg of ferric nitrate. As soon as the blue color of the mixture has disappeared, 1.1 g of metallic sodium are added over 15 minutes. The mixture is stirred at -33 ° C for 2 hours, a solution of 3.6 g of cyclosporin A in 60 cm ³ of tetrahydrofuran is added dropwise in approximately 20 minutes and then a solution of 3.1 g of di disulfide - [2- (N-methyl-Ni.propylamino) ethyl] in 15 cm ³ of tetrahydrofuran is added in 15 minutes. The reaction mixture is stirred at a temperature in the region of -33 ° C for 1 hour, then 3 g of solid ammonium chloride are added in portions. With stirring, the ammonia is allowed to evaporate by passing the temperature of the mixture from -33 to 25 ° C in 12 hours. The mixture is diluted with 100 cm ³ of diethyl ether and then filtered. The solid is rinsed with 100 cm ³ in total of diethyl ether. The combined organic phases are concentrated under reduced pressure (2.7 kPa), at a temperature in the region of 40 ° C. The solid obtained (6.3 g) is purified by chromatography on a silica column (0.020-0.045 mm) (eluent: ethyl acetate-methanol 4: 1 by volume) and collecting 50 cm ³ fractions. The fractions containing the expected product (fractions 32 to 48) are combined and concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to give 0.640 g of a colorless lacquer which, treated with 30 cm ³ d distilled water gives, after filtration and drying at a temperature in the region of 40 ° C, 0.390 g of [(R) -2- (N-methyl-Ni.propylamino) ethylthio-Sar] ³ cyclosporin A in the form of a solid off-white melting around 70 ° C.

¹ H NMR spectrum (400 MHz, CDCl ₃ , δ in ppm): 1.26 (d, J = 7.5 Hz, 3H: CH ₃ 8β); 1.37 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 2.18 (s, 3H: NCH ₃ of 2 N-methyl N-isopropyl aminoethylthio in 3α); from 2.55 to 2.75 (mt: 4H corresponding to SCH ₂ CH ₂ N of 2 N-methyl N-isopropyl aminoethylthio in 3α); 2.71 - 2.72 - 3.12 - 3.14 - 3.27 - 3.45 and 3.51 (7 s, 3H each: the 7 NCH ₃ ); 2.83 (mt, 1H: NCH of 2 N-methyl N-isopropyl aminoethylthio in 3α); 3.65 (d, J = 6 Hz, 1H: OH in 1β); 3.77 (mt, 1H: CH 1β); 4.54 (mt, 1H: CH 7α); 4.65 (broad t, J = 9 Hz, 1H: CH 5α); 4.84 (mt, 1H: CH 8α); 4.97 (dd, J = 10.5 and 6 Hz, 1H: CH α of a leucine); 5.00 to 5.10 (mt, 2H: CH α of a leucine and CH 2α); 5.13 (d, J = 11 Hz, 1H: CH 11α); 5.24 (dd, J = 11.5 and 4 Hz, 1H: CH α of a leucine); 5.33 (mt, 2H: CH = CH); 5.50 (d, J = 6 Hz, 1H: CH 1α); 5.71 (dd, J = 10.5 and 4 Hz, 1H: CH α of a leucine); 5.97 (s, 1H: CH 3α); 7.17 (d, J = 8 Hz, 1H: CONH in 8); 7.34 (d, J = 9 Hz, 1H: CONH in 5); 7.66 (d, J = 8 Hz, 1H: CONH at 7); 7.95 (d, J = 10 Hz, 1H: CONH in 2).

Di [2- (N-methyl-N-i.propylamino) ethyl disulfide can be prepared as follows:

To a solution of 20 g of 2- (Ni.propyl-N-methylamino) -ethanethiol in 150 cm ³ of diethyl ether, 60 cm ³ of a 5N aqueous solution of sodium hydroxide are added and then passed an air current for 12 hours at a temperature in the region of 20 ° C. The mixture is extracted using 150 cm ³ in total of diethyl ether. The combined organic phases are washed with 100 cm ³ of distilled water, dried over magnesium sulphate and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C, to yield 11.1 g of disulphide di- [2- (N-methyl-Ni.propylamino) ethyl] in the form of a colorless oil.

2- (N-isopropyl-N-methylamino) -ethanethiol is prepared according to the following method:

A solution of 115 cm ³ of Ni.propyl-N-methylamine and 44 cm ³ of ethylene episulfide in 400 cm ³ of diethyl ether is heated to a temperature close to reflux for 36 hours. A fractional distillation under reduced pressure (2.5 kPa) of the reaction mixture leads to 43 g of 2- (Ni.propyl-N-methylamino) -ethanethiol in the form of a colorless oil boiling at 60 ° C under 2.5 kPa.

Example 5

[(R) -2- (N-methyl-Nt.butylamino) ethylthio-Sar] ³ cyclosporin A is prepared according to the following method:

To 100 cm ³ of ammonia maintained at a temperature in the region of -33 ° C., 100 mg of metallic sodium is added, then 100 mg of ferric nitrate. As soon as the blue color of the mixture has disappeared, 1.0 g of metallic sodium is added over 30 minutes. The mixture is stirred at -33 ° C for 1 hour, a solution of 3.6 g of cyclosporin A in 50 cm ³ of tetrahydrofuran is added dropwise in approximately 30 minutes then a solution of 3.5 g of di disulfide - [2- (N-methyl-Nt.butylamino) ethylel in 15 cm ³ of tetrahydrofuran is added in 15 minutes. The reaction mixture is stirred at a temperature in the region of -33 ° C for 1 hour, then 3 g of solid ammonium chloride are added in portions. With stirring, the ammonia is allowed to evaporate by passing the temperature of the mixture from -33 to 25 ° C in 12 hours. The mixture is diluted with 100 cm ³ of diethyl ether and then filtered. The solid is rinsed with 300 cm ³ in total of diethyl ether. The combined organic phases are concentrated under reduced pressure (2.7 kPa), at a temperature in the region of 40 ° C. The solid obtained is triturated with 250 cm ³ of pentane and then filtered. The residual solid (6 g) is purified by chromatography on a silica column (0.020-0.045 mm) (eluent: ethyl acetate-methanol 4: 1 by volume) and collecting 100 cm ³ fractions. The fraction containing the expected product (fraction 9) is concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to give 0.500 g of a solid which is stirred with 30 cm ³ of pentane and gives, after filtration and drying at a temperature in the region of 40 ° C., 0.200 g of [(R) -2- (N-methyl-Nt-butylamino) ethylthio-Sar] ³ cyclosporin A in the form of an off-white solid melting towards 130 ° C.

¹ H NMR spectrum (400 MHz, CDCl ₃ , δ in ppm): 1.05 (s: the 9 H corresponding to C (CH ₃ ) ₃ ); 1.26 (d, J = 7.5 Hz, 3H: CH ₃ 8β); 1.36 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 1.63 (d, J = 5 Hz, 3H: CH ₃ 1η); 2.19 (s, 3H: NCH ₃ of 2 N-terbutyl N-methyl aminoethylthio in 3α); 2.55 to 2.80 (mt: 4H corresponding to SCH ₂ CH ₂ N of 2 N-terbutyl N-methyl aminoethylthio in 3α); 2.70 - 2.72 - 3.12 - 3.13 - 3.26 - 3.45 and 3.51 (7 s, 3H each: the 7 NCH ₃ ); 3.61 (d, J = 6.5 Hz, 1H: OH in 1β); 3.76 (mt, 1H: CH 1β); 4.54 (mt, 1H: CH 7α); 4.65 (broad t, J = 9 Hz, 1H: CH 5α); 4.84 (mt, 1H: CH 8α); 4.97 (dd, J = 10 and 6 Hz, 1H: CH α of a leucine); 5.00 to 5.10 (mt, 2H: CH α of a leucine and CH 2α); 5.13 (d, J = 11 Hz, 1H: CH 11α); 5.24 (dd, J = 10.5 and 4 Hz, 1H: CH α of a leucine); 5.34 (mt, 2H: CH = CH); 5.49 (d, J = 6 Hz, 1H: CH 1α); 5.71 (dd, J = 10.5 and 4 Hz, 1H: CH α of a leucine); 5.90 (s, 1H: CH 3α); 7.16 (d, J = 8 Hz, 1H: CONH in 8); 7.33 (d, J = 9 Hz, 1H: CONH in 5); 7.66 (d, J = 8 Hz, 1H: CONH at 7); 7.96 (d, J = 10 Hz, 1H: CONH in 2).

Di- [2- (N-methyl-N-t.butylamino) ethyl disulfide] can be prepared as follows:

To a solution of 28.7 g of 2- (Nt.butyl-N-methylamino) -ethanethiol in 190 cm ³ of methanol, 0.1 cm ³ of a 1N aqueous solution of sodium hydroxide is added and then made pass a current of air for 60 hours at a temperature in the region of 20 ° C. The methanol is removed under reduced pressure (2.7 kPa). The residual oil is dissolved in 400 cm ³ of diethyl ether. The organic phase is dried over magnesium sulfate, filtered and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C, to yield 26.6 g of di- [2- (N-methyl) disulfide -Nt.butylamino) ethyl] in the form of a yellow oil.

2- (N-t.butyl-N-methylamino) -ethanethiol can be prepared according to the following method:

A solution of 125 cm ³ of Nt.butyl-N-methylamine and 50 g of ethylene episulfide in 750 cm ³ of t.butyl methyl ether is stirred for 48 hours at a temperature close to reflux. The mixture is concentrated under reduced pressure (10 kPa) at a temperature in the region of 35 ° C. A fractional distillation under reduced pressure (5.8 kPa) of the reaction mixture leads to 28.7 g of 2- (Nt.butyl-N-methylamino) -ethanethiol in the form of a colorless oil boiling between 84 and 86 ° C at 5.8 kPa.

Example 6

[(R) -2- (1-imidazolyl) -ethylthio-Sar] ³ cyclosporine A is prepared according to the following method:

To 80 cm ³ of ammonia maintained at a temperature in the region of -33 ° C., 100 mg of metallic sodium is added, then 100 mg of ferric nitrate.
As soon as the blue color of the mixture has disappeared, 0.82 g of metallic sodium is added over 15 minutes. The mixture is stirred at -33 ° C for 15 minutes, then a solution of 2.4 g of cyclosporin A in 10 cm ³ of tetrahydrofuran is added dropwise in approximately 15 minutes and then 5 g of di- [2- (1-imidazolyl) ethyl] solid are added in portions over 15 minutes.
The reaction mixture is stirred at a temperature in the region of -33 ° C for 3 hours, then 3.4 g of solid ammonium chloride are added in portions. With stirring, the ammonia is allowed to evaporate by passing the temperature of the mixture from -33 to 25 ° C in 12 hours. The mixture is diluted with 100 cm ³ of distilled water. The organic phase is decanted, the aqueous phase is washed with 3 times 50 cm ³ of ethyl acetate. The combined organic phases are dried over magnesium sulfate, filtered and then concentrated under reduced pressure (2.7 kPa), at a temperature in the region of 40 ° C. The beige meringue thus obtained (2.25 g) is purified by chromatography on a column of silica (0.020-0.045 mm) (eluent: ethyl acetate-methanol 19: 1 by volume) and collecting 20 cm ³ fractions .
The fractions containing the expected product are concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to give 0.520 g of a solid. This solid, triturated with 50 cm ³ of pentane gives, after filtration and drying at a temperature in the region of 40 ° C, 0.420 g of a solid which is purified by chromatography on a second column of silica (0.020-0.045 mm) (eluent: ethyl acetate-methanol 4: 1 by volume). The fractions containing the expected product are concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The residual solid is stirred with 10 cm ³ of pentane to give, after filtration and drying at a temperature in the region of 40 ° C, 0.245 g of [(R) -2- (1-imidazolyl) -ethylthio-Sar] ³ cyclosporine A in the form of a yellow solid, melting at around 208 ° C.

¹ H NMR spectrum (400 MHz, CDCl ₃ , at a temperature of 333 K, δ in ppm): 1.26 (d, J = 7.5 Hz, 3H: CH ₃ 8β); 1.36 (d, J = 7.5 Hz, 3H: CH ₃ 7β); 1.63 (d, J = 5 Hz, 3H: CH ₃ 1η); 2.72 - 3.07 - 3.15 - 3.25 - 3.40 and 3.50 (6 s, respectively 6H - 3H - 3H - 3H - 3H - 3H: the 7 NCH ₃ ); from 3.80 to 3.95 (mt, 2H: CH 1β and OH in 1β); 4.16 (mt, 2H: NCH ₂ ); 4.51 (mt, 1H: CH 7α); 4.71 (broad t, J = 9 Hz, 1H: CH 5α); 4.86 (mt, 1H: CH 8α); from 4.95 to 5.10 (mt, 3H: CH α of two leucines and CH 2α); 5.17 (d, J = 11 Hz, 1H: CH 11α); 5.22 (dd, J = 11.5 and 4 Hz, 1H: CH α of a leucine); 5.35 (mt, 2H: CH = CH); 5.42 (d, J = 6 Hz, 1H: CH 1α); 5.72 (dd, J = 10.5 and 4 Hz, 1H: CH α of a leucine); 5.82 (s, 1H: CH 3α); 6.93 - 7.10 and 7.54 (3 seconds wide, 1 hour each: aromatic imidazole H); 7.12 (d, J = 8 Hz, 1H: CONH at 8); 7.19 (d, J = 9 Hz, 1H: CONH in 5); 7.53 (mt, 1H: CONH in 7); 7.87 (d, J = 10 Hz, 1H: CONH in 2).

Di- [2- (1-imidazolyl) ethyl] disulfide can be prepared from as follows:

To a solution of 15 g of 2- (1-imidazolyl) -ethanethiol in 200 cm ³ of dichloromethane cooled to 0 ° C., 32.3 cm ³ of triethylamine is added dropwise over 10 minutes, then a solution of 14, 59 g of iodine in 68 cm ³ of diethyl ether. The mixture is stirred for 30 minutes at a temperature in the region of 20 ° C and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The pasty residue is triturated in the presence of 50 cm ³ of isopropanol. The solid formed is filtered and rinsed with 25 cm ³ in total of isopropanol to give a first jet of di- [2- (1-imidazolyl) ethyl disulfide]. The combined organic phases are concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C.

The pasty residue is triturated in the presence of 50 cm ³ of ethyl acetate. The solid formed is filtered to give a second jet of di- [2- (1-imidazolyl) ethyl disulfide]. The two jets of di- [2- (1-imidazolyl) ethyl] disulfide are combined and dried under vacuum (10 kPa) at a temperature in the region of 20 ° C. to yield 14.2 g of di- [2 disulfide - (1-imidazolyl) ethyl].

2- (1-imidazolyl) -ethanethiol can be prepared in the following manner next:

A solution of 28.34 g of 2- (1-imidazolyl) ethylisothiourea hydrochloride and 18.56 g of sodium hydroxide in 300 cm ³ of distilled water is heated at reflux for 150 minutes. After having been brought to a temperature in the region of 20 ° C., the mixture is acidified by adding concentrated hydrochloric acid (20 cm ³ ) and then brought to pH = 7 by adding a saturated solution of sodium bicarbonate. The mixture is extracted with 600 cm ³ in total of ethyl acetate. The combined organic phases are washed with 100 cm ³ in total of distilled water, filtered, dried over magnesium sulfate and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C to yield 15.0 g of 2- (1-imidazolyl) -ethanethiol in the form of a yellow oil.

2- (1-imidazolyl) ethylisothiourea hydrochloride can be prepared as follows:

To a solution of 30 g of 2-hydroxy- (1-imidazolyl) -ethane in 300 cm ³ of dichloromethane is added dropwise in 30 minutes 44.2 cm ³ of thionyl chloride and then the mixture is brought to a neighboring temperature reflux for 16 hours. The mixture is concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The pasty residue is treated with 100 cm ³ of dichloromethane and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. This step is repeated twice. A suspension of 34.9 g of crude 2-chloro- (1-imidazolyl) -ethane hydrochloride thus obtained and 15.93 g of thiourea in 125 cm ³ of dimethylformamide is heated to a temperature in the region of 110 ° C for 90 minutes .

The mixture is cooled to a temperature in the region of 20 ° C. The yellow solid formed is then filtered, rinsed with 100 cm ³ in total of diethyl ether and dried under vacuum (10 kPa) at a temperature in the region of 40 ° C to yield 28.34 g of 2- hydrochloride (1 -imidazolyl) ethylisothiourea in the form of a solid melting at a temperature in the region of 206 ° C.

2-hydroxy- (1-imidazolyl) -ethane can be prepared in the following manner next:

To a suspension of 30 g of sodium hydride (50% in mineral oil) in 250 cm ³ of dimethylformamide is added in 30 minutes a solution of 68 g of imidazole of 250 cm ³ of dimethylformamide. The mixture is stirred for 90 minutes at a temperature in the region of 20 ° C. and then a solution of 50.5 g of 2-chloroethanol in 50 cm ³ of dimethylformamide is added in one hour. The mixture is stirred for 12 hours at a temperature in the region of 20 ° C. and then filtered. The filtrate is treated with 100 cm ³ of distilled water and then concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 55 ° C. The pasty residue is taken up in 150 cm ³ of petroleum ether, the liquid phase is decanted and the residue is triturated for one hour with 100 cm ³ of isopropanol. The precipitate formed is filtered, the filtrate is concentrated under reduced pressure (2.7 kPa) at a temperature in the region of 40 ° C. The residual oil (113.1 g) is distilled under reduced pressure (5 kPa) to yield 105.7 g of 2-hydroxy- (1-imidazolyl) -ethane in the form of a yellow oil distilling at a temperature from 180-183 ° C at 5 kPa.

Example 7

[(R)-2-aminoéthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-méthylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-éthylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-i.propylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-t.butylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-phénylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-benzylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-méthyl-N-éthylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-méthyl-N-allylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-méthyl-N-phénylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N-méthyl-N-benzylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N,N-di-i.propylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(N,N-diallylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-3-aminopropylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-éthylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-i.propylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-t.butylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-phénylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-benzylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthyl-N-éthylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthyl-N-i.propyllamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthyl-N-t.butylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthyl-N-allylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthyl-N-phénylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N-méthyl-N-benzylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-2-(N,N-diméthylamino)éthylthio-Sar]³-cyclosporine A ;

[(R)-3-(N,N-diéthylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N,N-di-i.propylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(N,N-diallylamino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(1-pipéridyl)propylthio-Sar]³-cyclosporine A ;

[(R)-4-aminobutylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-éthylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-i.propylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-t.butylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-phénylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-benzylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthyl-N-éthylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthyl-N-i.propylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthyl-N-t.butylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthyl-N-allylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthyl-N-phénylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N-méthyl-N-benzylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N,N-diméthylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N,N-diéthylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N,N-di-i.propylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(N,N-diallylamino)butylthio-Sar]³-cyclosporine A ;

[(R)-4-(1-pipéridyl)butylthio-Sar]³-cyclosporine A ;

[(R)-2-amino-2-méthylpropylthio-Sar]³-cyclosporine A ;

[(R)-2-(N,N-diméthylamino)-2-méthylpropylthio-Sar]³-cyclosporine A ;

[(R)-2-(N,N-diéthylamino)-2-méthylpropylthio-Sar]³-cyclosporine A ;

[(R)-2-(1-pipéridyl)-2-méthylpropylthio-Sar]³-cyclosporine A ;

[(R)-3-amino-3-méthylbutylthio-Sar]³-cyclosporine A ;

[(R)-3-(N,N-diméthylamino)-3-méthylbutylthio-Sar]³-cyclosporine A ;

[(R)-3-(N,N-diéthylamino)-3-méthylbutylthio-Sar]³-cyclosporine A ;

[(R)-3-(1-pipéridyl)-3-méthylbutylthio-Sar]³-cyclosporine A ;

[(R)-2-(1-morpholino)éthylthio-Sar]³-cyclosporine A ;

[(R)-2-(1-azétidino)éthylthio-Sar]³-cyclosporine A ;

{(R)-2-[1-(4-méthylpipérazino)}éthylthio-Sar}³-cyclosporine A ;

{(R)-2-[1-(4-phénylpipérazino)]éthylthio-Sar}³-cyclosporine A ;

{(R)-2-[1-(4-benzylpipérazino)]éthylthio-Sar}³-cyclosporine A ;

{(R)-2-[1-(4-méthyl-1,2,3,6-tétrahydropyridyl)]éthylthio-Sar}³-cyclosporine A ;

{(R)-2-[1-(4-phényl-1,2,3,6-tétrahydropyridyl)]éthylthio-Sar}³-cyclosporine A ;

[(R)-3-(1-morpholino)propylthio-Sar]³-cyclosporine A ;

[(R)-3-(1-azétidino)propylthio-Sar]³-cyclosporine A ;

{(R)-3-[1-(4-méthylpipérazino)]propylthio-Sar}³-cyclosporine A ;

{(R)-3-[1-(4-phénylpipérazino)]propylthio-Sar}³-cyclosporine A ;

{(R)-3-[1-(4-benzylpipérazino)]propylthio-Sar}³-cyclosporine A ;

{(R)-3-[1-(4-méthyl-1,2,3,6-tétrahydropyridyl)]propylthio-Sar}³-cyclosporine A ;

{(R)-3-[1-(4-méthyl-1,2,3,6 tétrahydropyridyl)]propylthio-Sar}³-cyclosporine A.

By following a procedure analogous to the methods described in Examples 1 to 3, the following products are prepared:

[(R) -2-aminoethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-methylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-ethylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (Ni.propylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (Nt.butylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-phenylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-benzylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-methyl-N-ethylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-methyl-N-allylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-methyl-N-phenylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N-methyl-N-benzylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N, N-di-i.propylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N, N-diallylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -3-aminopropylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-ethylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (Ni.propylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (Nt.butylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-phenylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-benzylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methyl-N-ethylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methyl-Ni.propyllamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methyl-Nt.butylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methyl-N-allylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methyl-N-phenylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N-methyl-N-benzylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N, N-diethylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N, N-di-i.propylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N, N-diallylamino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (1-piperidyl) propylthio-Sar] ³ -cyclosporin A;

[(R) -4-aminobutylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-ethylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (Ni.propylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (Nt.butylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-phenylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-benzylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methyl-N-ethylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methyl-Ni.propylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methyl-Nt.butylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methyl-N-allylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methyl-N-phenylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N-methyl-N-benzylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N, N-dimethylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N, N-diethylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N, N-di-i.propylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (N, N-diallylamino) butylthio-Sar] ³ -cyclosporin A;

[(R) -4- (1-piperidyl) butylthio-Sar] ³ -cyclosporin A;

[(R) -2-amino-2-methylpropylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N, N-dimethylamino) -2-methylpropylthio-Sar] ³ -cyclosporin A;

[(R) -2- (N, N-diethylamino) -2-methylpropylthio-Sar] ³ -cyclosporin A;

[(R) -2- (1-piperidyl) -2-methylpropylthio-Sar] ³ -cyclosporin A;

[(R) -3-amino-3-methylbutylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N, N-dimethylamino) -3-methylbutylthio-Sar] ³ -cyclosporin A;

[(R) -3- (N, N-diethylamino) -3-methylbutylthio-Sar] ³ -cyclosporin A;

[(R) -3- (1-piperidyl) -3-methylbutylthio-Sar] ³ -cyclosporin A;

[(R) -2- (1-morpholino) ethylthio-Sar] ³ -cyclosporin A;

[(R) -2- (1-azetidino) ethylthio-Sar] ³ -cyclosporin A;

{(R) -2- [1- (4-methylpiperazino)} ethylthio-Sar} ³ -cyclosporin A;

{(R) -2- [1- (4-phenylpiperazino)] ethylthio-Sar} ³ -cyclosporin A;

{(R) -2- [1- (4-benzylpiperazino)] ethylthio-Sar} ³ -cyclosporin A;

{(R) -2- [1- (4-methyl-1,2,3,6-tetrahydropyridyl)] ethylthio-Sar} ³ -cyclosporin A;

{(R) -2- [1- (4-phenyl-1,2,3,6-tetrahydropyridyl)] ethylthio-Sar} ³ -cyclosporin A;

[(R) -3- (1-morpholino) propylthio-Sar] ³ -cyclosporin A;

[(R) -3- (1-azetidino) propylthio-Sar] ³ -cyclosporin A;

{(R) -3- [1- (4-methylpiperazino)] propylthio-Sar} ³ -cyclosporin A;

{(R) -3- [1- (4-phenylpiperazino)] propylthio-Sar} ³ -cyclosporin A;

{(R) -3- [1- (4-benzylpiperazino)] propylthio-Sar} ³ -cyclosporin A;

{(R) -3- [1- (4-methyl-1,2,3,6-tetrahydropyridyl)] propylthio-Sar} ³ -cyclosporin A;

{(R) -3- [1- (4-methyl-1,2,3,6 tetrahydropyridyl)] propylthio-Sar} ³ -cyclosporin A.

The present invention also relates to pharmaceutical compositions containing at least one product of general formula (I) the case if necessary in the form of salt, in the pure state or in the form of a combination with one or more compatible diluents or adjuvants and pharmaceutically acceptable, or with another agent antiretrovirus, possibly intended for the treatment of AIDS, a antiviral, immunomodulatory or antimicrobial agent.

The composition according to the invention is capable of keeping alive the cells infected with a retrovirus such as for example HIV and therefore reduce the progression to AIDS or decrease its severity in subjects already infected by reducing mortality from infected cells. The compositions can be used by the oral, parenteral, rectal or aerosol.

The pharmaceutical compositions can be used as curative or preventive in subjects with immunodeficiency and / or infected with a retrovirus. Of course, the constitution of these compositions will be adapted to the particular case of digestive tract of the immunocompromised.

As solid compositions for oral administration may be used tablets, pills, capsules, powders or granules. In these compositions, the active product according to the invention is mixed with one or more inert diluents or adjuvants, such than sucrose, lactose or starch.

These compositions can include substances other than thinners, for example a lubricant such as magnesium stearate or a coating for controlled release.

As liquid compositions for oral administration, it is possible to use pharmaceutically acceptable solutions, suspensions, emulsions, syrups and elixirs containing inert diluents such as water or paraffin oil. These compositions may also include substances other than thinners, for example wetting, sweetening or flavorings.

The compositions for parenteral administration can be sterile solutions or emulsions. As solvent or vehicle, can use propylene glycol, polyethylene glycol, oils vegetable, in particular olive oil, organic esters injectables, for example ethyl oleate.

These compositions can also contain adjuvants, in particular particularly wetting, isotonizing, emulsifying agents, dispersants and stabilizers.

Sterilization can be done in several ways, for example using a bacteriological filter, by irradiation or by heating. They can also be prepared in the form of compositions sterile solids which can be dissolved at the time of use in sterile water or any other sterile injectable mixture.

The compositions for rectal administration are the suppositories or rectal capsules, which contain in addition to the active ingredient, excipients such as cocoa butter, semi-synthetic glycerides or polyethylene glycols.

The compositions can also be aerosols. For use in the form of liquid aerosols, the compositions can be stable sterile solutions or solid compositions dissolved in time of use in sterile pyrogen-free water, in serum or any other pharmaceutically acceptable vehicle. For use under form of dry aerosols intended to be directly inhaled, the active ingredient is finely divided and associated with a diluent or solid water-soluble vehicle with a particle size of 30 to 80 µm, by for example dextran, mannitol or lactose.

In human therapy, the doctor will determine the dosage he considers the most appropriate based on preventive treatment or curative, depending on age, weight, degree of infection and other factors specific to the subject at hand. Generally, doses are between 5 and 30 mg / kg orally for a adult.

It has also been shown that the cyclosporine derivatives of formula general (I) show a synergistic effect or at least of addition when combined with other antiviral agents active on retroviruses. The present invention also relates to synergizing associations which contain at least one derivative of cyclosporine of general formula (I) and / or, where appropriate, their salts, and an active ingredient known for its activity on retrovirus.

Agents known for their activity on retroviruses that can be associated, are chosen from compatible and inert agents with respect to the cyclosporine derivative of general formula (I), also both in the category of pharmacological treatments than in the category of alternative treatments such as gene therapy and cellular or antisense. Without limitation these agents constituting the different therapeutic classes are chosen by example among nucleoside inhibitors (NRTI) and not nucleoside (NNRTI) of reverse transcriptase [zidovudine (AZT), didanosine (DDI), dideoxycytidine (DDC), d4T, ribavirin, 3TC, nevirapine ...], among protease inhibitors [such as for example Saquinavir, Ritonavir, Indinavir and Nelfinavir], integrase inhibitors [such as AR177], among gene therapy inhibitors targeting regulatory proteins replication of HIVs such as protein inhibitors rev [such as Rev M10], or nucleocapsid [such as DIBAs], among the inhibitors targeting the messenger RNA transcripts specific for all HIVs like for example anti-sense [like GEM92, GPI-2A ...], among the inhibitors of the cellular dNTP modulator family [such as hydroxyurea], among cytokine inhibitors [like TNF], among HIV entry inhibitors [such as T20, SPC-3 ...], as well as among the constituent agents the therapeutic classes used in vaccine approaches both by biotechnology [like HIVAC-le, ALVAC ...] only by compounds acting on the immune response [like RG-8394].

In particular, the cyclosporine derivative of Example 2 manifests a particularly interesting effect when combined with AZT, ddI, Sasquinavir, and ribavirin.

Pharmaceutical compositions comprising such combinations, optionally in the presence of pharmaceutical excipients acceptable, also fall within the scope of this invention.

The following example illustrates a composition according to the invention.

Example

A formulation which can be administered orally and which has the following composition is prepared: [(R) -2- (N, N-dimethylamino) ethylthio-Sar] ³ -cyclosporin A 250 mg Magnesium stearate 3 mg Acidsol 15 mg Colloidal silica 2 mg Lactose 130 mg

Claims (11)

1. Cyclosporin derivative, characterized in that it corresponds to the general formula :

   

   in which:

   Alk represents an alkylene radical containing 2 to 6 carbon atoms in straight or branched chains or a cycloalkylene radical containing 3 to 6 carbon atoms and

   R represents

   either a carboxyl or alkyloxycarbonyl radical,

   or an -NR₁R₂ radical in which R₁ and R₂, which are identical or different, represent hydrogen atoms or alkyl, alkenyl (2 to 4C), cycloalkyl (3 to 6C) or optionally substituted (by a halogen atom, alkyloxy, alkyloxycarbonyl, amino, alkylamino or dialkylamino) phenyl radicals or represent benzyl or heterocyclyl radicals, the heterocyclyl radical being saturated or unsaturated and containing 5 or 6 ring members and 1 to 3 heteroatoms, or in which R₁ and R₂ form, with the nitrogen atom to which they are attached, a saturated or unsaturated heterocycle containing 4 to 6 ring members, which can contain another heteroatom chosen from nitrogen, oxygen or sulphur, optionally substituted by alkyl, phenyl or benzyl,

   or a radical of general formula:

   

   in which R₁ and R₂ are defined as above, R₃ represents a hydrogen atom or an alkyl radical and n is an integer from 2 to 4,
   the alkyl portions or radicals defined above being straight or branched and containing 1 to 4 carbon atoms, and its pharmaceutically acceptable salts, when they exist.
2. Cyclosporin derivative according to Claim 1, characterized in that:

   Alk represents an alkylene radical containing 2 to 6 carbon atoms in straight or branched chains and

   R represents an -NR₁R₂ radical in which R₁ and R₂, which are identical or different, represent hydrogen atoms or alkyl, alkenyl (2 to 4C) or optionally substituted (by a halogen atom, alkyloxy, alkyloxycarbonyl, amino, alkylamino or dialkylamino) phenyl radicals or represent benzyl radicals or in which R₁ and R₂ form, with the nitrogen atom to which they are attached, a saturated or unsaturated heterocycle containing 4 to 6 ring members, which can contain another heteroatom chosen from nitrogen, oxygen or sulphur, optionally substituted by alkyl, the alkyl portions or radicals defined above being straight or branched and containing 1 to 4 carbon atoms, and its pharmaceutically acceptable salts, when they exist.
3. Cyclosporin derivative according to Claim 1, characterized in that:

   Alk represents an alkylene radical containing 2 to 5 carbon atoms in straight or branched chains and

   R represents an -NR₁R₂ radical in which R₁ and R₂, which are identical or different, represent hydrogen atoms or alkyl, allyl, phenyl or benzyl radicals or in which R₁ and R₂ form, with the nitrogen atom to which they are attached, a heterocycle chosen from azetidinyl, piperidyl, piperazinyl, N-methylpiperazinyl, N-phenylpiperazinyl, N-benzylpiperazinyl, imidazolyl, morpholino, tetrahydropyridyl, methyltetrahydropyridyl or phenyltetrahydropyridyl, the alkyl portions or radicals defined above being straight or branched and containing 1 to 4 carbon atoms, and its pharmaceutically acceptable salts, when they exist.
4. Cyclosporin derivative according to Claim 1, characterized in that it is [(R)-2-(N,N-diethylamino) ethylthio-Sar]³-cyclosporin A, and its pharmaceutically acceptable salts.
5. Cyclosporin derivative according to Claim 1, characterized in that it is [(R)-2-(N,N-dimethylamino)ethylthio-Sar]³-cyclosporin A, and its pharmaceutically acceptable salts.
6. Cyclosporin derivative according to Claim 1, characterized in that it is [(R)-2-(1-piperidyl)ethylthio-Sar]³-cyclosporin A, and its pharmaceutically acceptable salts.
7. Cyclosporin derivative according to Claim 1, characterized in that it is [(R)-2-(N-methyl-N-i-propylamino)ethylthio-Sar]³-cyclosporin A, and its pharmaceutically acceptable salts.
8. Cyclosporin derivative according to Claim 1,
   characterized in that it is [(R)-2-(N-methyl-N-t-butylamino)ethylthio-Sar]³-cyclosporin A, and its pharmaceutically acceptable salts.
9. Process for the preparation of Cyclosporin derivative according to Claim 1, characterized in that a disulphide of general formula: R-Alk-S-S-Alk-R in which R and Alk are defined as above, the functional groups of which can interfere with the reaction have, if appropriate, been protected beforehand, is reacted with an activated form of cyclosporin A, then, if appropriate, the protective radicals are removed and/or the product obtained is optionally converted into a salt, when they exist.
10. Pharmaceutical composition, characterized in that it contains at least one product according to Claim 1, in the pure state or in combination with any compatible and pharmaceutically acceptable diluent or adjuvant and/or in combination with another antiviral, immunomodulating or antimicrobial active principle.
11. Synergizing combinations, characterized in that they comprise at least one cyclosporin derivative according to Claim 1 and at least one other agent known for its anti-retrovirus activity.